US 11,702,649 B2
Single cell cellular component enrichment from barcoded sequencing libraries
Paul Blainey, Cambridge, MA (US); Navpreet Ranu, Cambridge, MA (US); Todd Gierahn, Cambridge, MA (US); and J. Christopher Love, Cambridge, MA (US)
Assigned to The Broad Institute, Inc., Cambridge, MA (US); and Massachusetts Institute of Technology, Cambridge, MA (US)
Appl. No. 16/758,640
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
PCT Filed Oct. 23, 2018, PCT No. PCT/US2018/057161
§ 371(c)(1), (2) Date Apr. 23, 2020,
PCT Pub. No. WO2019/084046, PCT Pub. Date May 2, 2019.
Claims priority of provisional application 62/576,031, filed on Oct. 23, 2017.
Claims priority of provisional application 62/736,863, filed on Sep. 26, 2018.
Prior Publication US 2020/0392479 A1, Dec. 17, 2020
Int. Cl. C12N 15/10 (2006.01); C12Q 1/6806 (2018.01); C12Q 1/6869 (2018.01); C40B 30/04 (2006.01); C40B 50/06 (2006.01); C12N 1/20 (2006.01); C40B 40/06 (2006.01); C40B 40/10 (2006.01); C12R 1/465 (2006.01)
CPC C12N 15/1006 (2013.01) [C12N 1/205 (2021.05); C12Q 1/6806 (2013.01); C12Q 1/6869 (2013.01); C40B 30/04 (2013.01); C40B 40/06 (2013.01); C40B 40/10 (2013.01); C40B 50/06 (2013.01); C12N 2310/20 (2017.05); C12R 2001/465 (2021.05); C12Y 301/11 (2013.01); C12Y 302/02027 (2013.01)] 18 Claims
OG exemplary drawing
 
1. A method of enriching barcoded constructs from a sequenced nucleic acid library generated from a plurality of cells comprising capturing nucleic acid library molecules from the sequenced nucleic acid library by targeting library transcripts comprising one or more cell-identifying barcodes
wherein the method is a method of identifying a transcriptome from at least one single cell or a subpopulation of single cells, said method comprising enriching library molecules from the at least one single cell or subpopulation of single cells based on the one or more unique cell-identifying barcodes, wherein the targeted barcodes identify transcripts of single cells represented within the sequenced nucleic acid library,
wherein targeting comprises introducing to the sequenced nucleic acid library an oligonucleotide, the oligonucleotide comprising a primer, a sequence for hybridizing to the one or more cell-identifying barcodes, and a label.